LOCUS       sequence-458385-        6737 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mtPyronicSF in neurons targeted to the mitochondria 
            serving as an optical sensor for mitochondrial pyruvate uptake. 
            Backbone from Addgene plasmid 100834.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6737)
  AUTHORS   Tiwari A, Myeong J, Hashemiaghdam A, Stunault MI, Zhang H, Niu X, 
            Laramie MA, Sponagel J, Shriver LP, Patti GJ, Klyachko VA, Ashrafi G
  TITLE     Mitochondrial pyruvate transport regulates presynaptic metabolism 
            and neurotransmission.
  JOURNAL   Sci Adv. 2024 Nov 15;10(46):eadp7423. doi: 10.1126/sciadv.adp7423. 
            Epub 2024 Nov 15.
  PUBMED    39546604
REFERENCE   2  (bases 1 to 6737)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2024-11-15"; volume: "10(46):eadp7423. doi"; pages: " 
            10.1126/sciadv.adp7423. Epub 2024 Nov 15"
COMMENT     Sequence Label: CamKII-mtPyronicSF
FEATURES             Location/Qualifiers
     source          1..6737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        149..1437
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             1470..1544
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     misc_feature    3096..3684
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3149..3169)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3567..3578)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3727..3848
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3738..3757)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3792..3811
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4000..4140
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4000..4129
                     /label=AAV2 ITR
     rep_origin      4215..4670
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4302..4321)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4512..4533
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4687..4706)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4806..4828
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4866..4884)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4952..5056
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5057..5125,5126..5917)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5275..5294)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6088..6676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6577..6596
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtttaaca ttatggcctt aggtcacttc atctccatgg
      181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc
      241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa
      301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag
      361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca
      421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt
      481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc
      541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca
      601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt
      661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg
      721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag
      781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt
      841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca
      901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg
      961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt
     1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg
     1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag
     1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa
     1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa
     1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc
     1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt
     1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct
     1441 agagcggtac cggatccacc cgccacccca tgtccgtcct gacgccgctg ctgctgcggg
     1501 gcttgacagg ctcggcccgg cggctcccag tgccgcgcgc caagatccat tcgttgctgc
     1561 aggtcgacct cgagatggca tatagcaaaa ttcggcagcc caaactgagc gatgtgattg
     1621 agcagcagct ggagtttctg attctggaag gcaccctgag acctggagag aaactgcccc
     1681 ctgagcgcga actcgccaag cagttcgacg tgagtcgacc atcactgagg gaggctatcc
     1741 agaggctgga agcaaaggga ctgctcctga ggagacaggg aggagggact ttcgtgcaga
     1801 gctccctgtg gcagagcttc agcgaccccc tggtcgagct cctgtctgac cacccagaaa
     1861 gtcagtacga tctcctggag acaagacatg ctctggaagg catcgccgct tactatgcag
     1921 ccctgcggtc cactgacgag gataaggaac gcatccgaga gctgcaccat gccattgaac
     1981 tcgctcagca gtcaggagac ctggatgcag agagcaacgc cgtgctgcag taccagattg
     2041 cagtcaccga ggctgcacac aatgtggtcc tcctgcatct cctgaggtgc atggagccaa
     2101 tgctggccca gaacgtgaga cagaattttg agctcctgct cgaaaacgtc tatatcaagg
     2161 ccgacaagca gaaaaacggc attaaggcta acttcaagat cagacacaac atcgaggatg
     2221 gtggcgtgca gctggcctac cattatcagc agaacacacc aatcggagac ggaccagtgc
     2281 tgctcccaga taatcactac ctgagcgtcc agtccaagct gtctaaagac cctaacgaga
     2341 agcgggatca tatggtgctg ctcgaatttg tcacagccgc tgggatcact ctgggtatgg
     2401 acgagctcta taaaggaggg accggtggca gtatggtgtc aaagggcgag gaactgttca
     2461 caggagtggt ccccattctg gtggagctcg acggcgatgt caatggacac aaattttccg
     2521 tgtctggcga gggcgaagga gatgctacct acgggaagct gacactcaaa ttcatctgca
     2581 ccacaggcaa gctgccagtg ccctggccta ctctggtcac taccctcacc tacggggtgc
     2641 agtgtttctc cagatatccc gaccacatga agcagcatga tttctttaaa tctgctatgc
     2701 ctgaggggta catccaggaa cggacaattt tctttaagga cgatggtaac tacaaaacac
     2761 gcgcagaggt gaagttcgaa ggcgacactc tggtcaatcg aatcgagctg aagggaattg
     2821 actttaaaga agatgggaac atcctgggtc acaagctgga gtacaatact aggtattctc
     2881 ggcgcgaaat gctgccactc gtgtctagtc acaggaccag aatctttgag gcaattatgg
     2941 ccggaaagcc cgaggaagct agagaagcaa gtcaccggca tctggccttc atcgaggaaa
     3001 ttctgctcga ccggagccgc gaggaatccc gaagggagcg cagcctgaga cgactcgaac
     3061 agcgaaagaa ctgactcctc gagaagctta tcgataatca acctctggat tacaaaattt
     3121 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3181 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3241 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3301 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3361 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3421 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3481 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     3541 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3601 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3661 tctccctttg ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg cttcgagcag
     3721 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     3781 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     3841 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     3901 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta
     3961 gagcatggct acgtaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt
     4021 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     4081 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     4141 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     4201 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4261 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     4321 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4381 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4441 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4501 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     4561 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     4621 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     4681 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4741 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4801 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4861 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4921 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4981 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5041 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5101 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5161 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5221 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5281 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5341 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5401 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5461 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5521 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5581 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5641 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5701 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5761 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5821 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5881 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5941 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6001 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6061 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6121 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6181 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6241 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6301 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6361 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6421 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6481 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6541 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6601 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6661 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6721 gccttttgct cacatgt
//
